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. Author manuscript; available in PMC: 2022 Aug 10.
Published in final edited form as: Nat Chem. 2022 Jun 20;14(8):905–913. doi: 10.1038/s41557-022-00972-7

Figure 6. Spatio-temporal control of translation, stability, and immune response of FlashCap-mRNAs in cells.

Figure 6

a) Irradiation of circled area in a confocal laser scanning microscope and analysis of fluorescence of HeLa cells transfected with FlashCap 2-eGFP-mRNA. Nuclei are stained by DAPI (blue). Scale bars are 100 µm or 50 µm (inset). Individual colour channels were adjusted. Shown is one representative image from n=3 independent experiments. The microscopy images were hyperstacked and the background subtracted (30 pixels) with ImageJ. b) Stability of FlashCap-mRNAs. RTqPCR data showing the relative RLuc mRNA level at 4 h or 10 h post transfection in HeLa cells. The 4 h time point is used for normalization and was set as 100 %. Data of n=3 independent experiments are shown as mean values +/- SEM. c) Immune response of FlashCap-mRNAs. FLuc activity of four different HEK-NF-ĸB cell lines (Null, TLR3, TLR7, TLR8) transfected with differently capped RLuc-mRNAs (either cap 0 or FlashCap1 or 2). TLR3, TLR7 and TLR8 indicates the overexpression of the respective Toll-Like-Receptor (TLR) in that cell line. Data are normalized to the cap 0-mRNA without irradiation. Data of n=4 independent experiments are shown as mean values +/- SEM.